The binding of Human SRY to DNA is an important step in human sex determination. Wild Type SRY binds to DNA and bends the DNA with an angle of approximately 70 degrees. Studies of human sex reversal have revealed that some single site mutations in SRY responsible for sex reversal demonstrate unusual behavior in terms of their relative binding affinity and alteration of the bending angle. Some mutants have been found with a large change in the binding affinity, and only a small change in the bending angle, and others have been found with a large change in the bending angle and a small change in the binding affinity. To better understand the mechanism of action of human SRY, these unusual mutants will be studied with ITC, in order to determine the changes in the energetics of binding. The bending angle of these mutants will be qualitatively analyzed using circular permutation assays. Any mutants exhibiting the conflicting behavior described above will be further analyzed using NMR to determine the structural factors that result in this strange behavior.